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SDS-page and western blot Cells (4 × 10 6 cells) were resuspended in 50 μl PBS buffer pH 7.2, mixed with the same volume of 2 × SDS/PAGE sample loading buffer (20 mM Tris–HCl, pH 8, 2 mM EDTA, 5% (m v −1 ) SDS, and 10% (v v −1 ) β-mercaptoethanol), and boiled for 5 min at 95°C